Research Article
Year 2017,
Volume: 48 Issue: 2, 73 - 79, 14.10.2017
Abstract
Carob (Ceratonia siliqua L. (Cesalpinaceae) is an evergreen tree. It naturally
grows in forests, but it is also commercially grown in some regions. As it was
in other plant species, molecular markers are used to identify genetic
diversity in carob. DNA isolation is the most significant factor effecting
molecular marker analysis. In this study, four different isolation protocols
were used to isolate quality DNA from young and old carob leaves. In young
leaves, while the lowest DNA concentrations were obtained from protocol A (5.73±0.89
ng/µl DNA), DNA concentration of Protocol D were
significantly higher than other protocols (900.66±23.38 ng/µl DNA).
In old leaves, the lowest DNA concentrations were obtained from protocol A (2.9±0.4
ng/µl DNA) and the highest DNA concentrations were obtained
from protocol C (11.36±0.8 ng/µl DNA). The DNA concentrations obtained
from the freshest tip leaves measured 656.66±36.95 ng/µl DNA.
DNA concentration of old leaves significantly lower than young leaves at all
protocols. While the lowest A260/280 nm absorbance ratios in young leaves
were observed in protocol A (0.76±0.02), the ratio in other protocols were between the
optimum values.
Keywords
References
- Akinnagbe O. M., Onah C. P., Olaolu, M. O., Ajayi, A. R., 2012. Farmers’ perception of the impact of climate change on agricultural activities in Otukpo local government area of Benue State, Nigeria. In: Proceedings of the 17th Annual National Conference AESON, 11 –14 March, 2012, pp109-117, Nsukka, Nigeria. Aubakirova, K., Omasheva, M., Ryabushkina, N.,Tazhibaev, T., Kampitova, G., Galiakparov, N., 2014. Evaluation of five protocols for DNA extraction from leaves of Malus siever, Vitis vinifera, and Armeniaca vulgaris. Genet. Mol. Res. 13 (1): 1278-1287. Ausubel, F. M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith. J.G. et al., 2002. Short Protocols in Molecular Biology: a compendium of methods from Current Protocols in Molecular Biyology, vol. 2,Wiley, New York, NY. Balik, J., Kyseláková, M., Vrchotová, N., Tríska, J., et al. 2008.Relations between polyphenols content and antioxidant activity in vine grapes and leaves. Czech J. Food Sci. 26: S25-S32 Barracosa, P., Almeida, M.T., Cenis, J., 1996. Characterization of cultivars of carob tree in Algarve (Portugal). In: Proceeding of the III International Carob Symposium. Cabanas-Tavira, Portugal Batlle, I.M., Tous, R.J., 1997. Carob germplasm characterization using izozymes. In: Proceeding of the III International Carob Symposium. Cabanas-Tavira, Portugal. Bushra, C., Afshan, Y., Tayyab, H., Riazuddin, S., 1999. Mini-scale genomic DNA extraction from cotton. Plant Mol. Biol. Rep., 17: 1–7. Carlson, W.A., 1986. carob: evaluation of trees, pods and kernels. Int. Tree Crops J., 3 (1986), pp. 281–290 Casıva, P.V., Saidman, B. O., Vılardı, J. C., Cıaldella, A.M., 2002. First comparative phonetic studies of Argentinean species of Acacia (Fabaceae), using morphometric, isozymal and RAPD approaches. AmericanJournal of Botany 89(5): 843–853. Crouch, H.K,. Crouch, J.H., Madsen, S., Vuylsteke, D.R., Ortız, R., 2000. Comparative analysis of phenotypic andgenotypic diversity among plantain landraces (Musa spp., AAB group). Theor. Appl. Genet, 101:1056–1065 Dellaporta, S.L., Wood, J., Hicks, J.B., 1983. A plant DNA mini preparation: Version II. Plant Mol. Bio. Rep. 1: 19-21. Doyle, J.J., Doyle, J.L., 1990. Isolation of plant DNA from fresh tissue. Focus 12: 13-15. Edwards, K., Johnstone, C.,Thompson. C., 1991. A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Res. 19: 1349. Emberger, L., Maire, R., 1941. Cataloguedes plantes du Maroc.Volume IV. Minerva, Alger. Garcia, M. G., . Ontivero, M., J. Dıazrıccı, C., Castagnaro, A., 2002. Morphological traits and high resolution RAPD markers for the identification of the main strawberry varieties cultivated in Argentina. Plant Breeding 121, 76-80. Gharnit, N., El Mtili, N., Ennabili, A., Sayah, F., 2004. Floral characterization of carob tree (Ceratonia siliqua L.) from the province of Chefchaouen (NW of Morocco). Moroccan J. Biol., 1pp. 41–5 Guillemaut, P., and Maréchal-Drouard, L., 1992. Isolation of plant DNA: a fast, inexpensive and reliable method. Plant Mol. Biol. Rep. 10: 60-65. Güzeldağ, G.,. Çolak, Õ., 2007. Molecular identification of Ganoderma lucidm from Turkey. Int. J. Agric. Biol., 9: 767–770. Halliwell, B., 1990. How to characterize a biological antioxidant. Free Radic. Res. Commun. 9: 1-32. Jobes, D.V., Hurley, D.L., Thien, L.B., 1995. Plant DNA isolation: a method to efficiently remove polyphenolics, polysaccharides and RNA. Taxon 44: 349-386. Konate, I., Berraho, E.B., Filali-Maltouf, A., 2009. Inter-simple sequence repeat markers variation among natural accessions of Moroccan Carob Tree (Ceratonia siliqua L.), Inter. J. of Agri.Biol., 11: 168-172. Konaté, I., Filali-Maltouf, A., Berraho, E., 2007. Diversity analysis of Moroccan carob (Ceratonia siliqua L.) accessions using phenotypic traits and RAPD markers, Acta Bot. Malacitana, 32: 79–90. Kulkarni M, Borse, T., Chaphalkar, S., 2001.Isolation and purification of genomic DNA from black plum (Eugenia jambolana Lam.) for analytical applications. Int. J. Biotechnol. Biochem. 3: 49-55. Lodhi, M.A., Ye, G.N., Weeden, N.F., Reisch, B.I., 1994. A simple and efficient method DNA extraction from grapevine caltivars and vitis species. Plant Mol. Biol. Rep., 12: 6–13. Morsy, A.A., 2007. Molecular variation of Achillea fragrantissima (Forssk.) SCH. BIP. Growing in five areas of South Sinai. Int. J. Agric. Biol., 9: 11–17 Ounzar, B., Hartmann, C., Rode, A., Benslimane, A., 1998. Date palm DNA minipreparation without liquid nitrogen. Plant Mol. Rep. 16: 263-269. Samal S., Rout, G.R., Lenkaet, P.C., -2003- Analysis of genetic relationships between populations of cashew (Anacardium occidentale L.) by using morphological characterisation and RAPD markers. Plant Soil Environ. 49, (4): 176-182. Samarakoon, T., Wang, S.Y., Alford, M.H., 2013. Enhancing PCR amplification of DNA from recalcitrant plant specimens using a trehalose-based additive. Appl. Plant Sci. 1: 1200236. Sytsma, K., Givnish, T.J., Simt, J.F., Hahn, W.J., 1993. Collection and Storage of Land Plant Samples for Macromolecular Comparisons. In: Methods in Enzymology - Molecular Evolution: Producing the Biochemical Data (Zimmer EA, White TJ, Cann RL and Wilson AC, eds.). Academic Press, San Diego, 23-38. Talhinhas, P., Neves-Martins, J., Leilâo, J., 2003. AFLP, ISSR and RAPD markers reveal high levels of genetic diversity among Lupinus spp. Plant Breed., 122: 507–510. Tous, J., Batlle, I., 1990. In: El Algarrobo (Ed). Mundi-Prensa, Madrid. Tous,J., 1992. Isozyme polymorphism in carob cultivars Hort. Sci., 27 (1992), pp. 257–258 Weishing, K., Nybom, H., Wolff, K., Meyer, W., 1995. DNA isolation and purification. In: DNA fingerprinting in plants and fungi, pp 44-59. CRC Press, Boca Raton, Florida.
Keçiboynuzunun Genç ve Olgun Yapraklarından DNA Ekstraksiyonu İçin Dört Farklı Protokolün Değerlendirilmesi
Abstract
Keçiboynuzu (Ceratonia siliqua L. (Cesalpinaceae) herdem yeşil bir bitkidir.
Keçiboynuzu doğal olarak ormanda yetişirken bazı bölgelerde ticari olarak da
yetiştirilmektedir. Diğer bitki türlerinde olduğu gibi moleküler markırlar
keçiboynuzunda da genetic çeşitliliği tanımlamak için kullanılmaktadır. Bu
çalışmada, keçiboynozunun genç ve olgun yapraklarında dört farklı protocol
kullanılarak kaliteli DNA elde etmek amaçlanmıştır. Genç yapraklarda, En düşük DNA
konsantrasyonu protocol A’dan (5.73±0.89 ng/µl DNA) sağlanırken, protocol D’nin (900.66±23.38
ng/µl DNA)
DNA konsantrasyonu diğer protokollerden oldukça yüksektir. Yaşlı yapraklarda,
en düşük DNA miktarı protokol A(2.9±0.4 ng/µl DNA)
ile en yüksek DNA miktarı protokol C(11.36±0.8 ng/µl DNA) şeklindedir. Genç yapraklardan elde
edilen DNA konsantrasyonu 656.66±36.95 ng/µl DNA
olarak ölçülmüştür. Olgun yapraklardaki DNA konsantrasyonları genç
yapraklardaki bütün protokollere göre oldukça düşüktür. Genç yapraklarda en
düşük emilim A260/280
nm ile protokol A (0.76±0.02) iken, diğer protokollerdeki oranlar optimum değerler
arasındadır.
Keywords
References
- Akinnagbe O. M., Onah C. P., Olaolu, M. O., Ajayi, A. R., 2012. Farmers’ perception of the impact of climate change on agricultural activities in Otukpo local government area of Benue State, Nigeria. In: Proceedings of the 17th Annual National Conference AESON, 11 –14 March, 2012, pp109-117, Nsukka, Nigeria. Aubakirova, K., Omasheva, M., Ryabushkina, N.,Tazhibaev, T., Kampitova, G., Galiakparov, N., 2014. Evaluation of five protocols for DNA extraction from leaves of Malus siever, Vitis vinifera, and Armeniaca vulgaris. Genet. Mol. Res. 13 (1): 1278-1287. Ausubel, F. M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith. J.G. et al., 2002. Short Protocols in Molecular Biology: a compendium of methods from Current Protocols in Molecular Biyology, vol. 2,Wiley, New York, NY. Balik, J., Kyseláková, M., Vrchotová, N., Tríska, J., et al. 2008.Relations between polyphenols content and antioxidant activity in vine grapes and leaves. Czech J. Food Sci. 26: S25-S32 Barracosa, P., Almeida, M.T., Cenis, J., 1996. Characterization of cultivars of carob tree in Algarve (Portugal). In: Proceeding of the III International Carob Symposium. Cabanas-Tavira, Portugal Batlle, I.M., Tous, R.J., 1997. Carob germplasm characterization using izozymes. In: Proceeding of the III International Carob Symposium. Cabanas-Tavira, Portugal. Bushra, C., Afshan, Y., Tayyab, H., Riazuddin, S., 1999. Mini-scale genomic DNA extraction from cotton. Plant Mol. Biol. Rep., 17: 1–7. Carlson, W.A., 1986. carob: evaluation of trees, pods and kernels. Int. Tree Crops J., 3 (1986), pp. 281–290 Casıva, P.V., Saidman, B. O., Vılardı, J. C., Cıaldella, A.M., 2002. First comparative phonetic studies of Argentinean species of Acacia (Fabaceae), using morphometric, isozymal and RAPD approaches. AmericanJournal of Botany 89(5): 843–853. Crouch, H.K,. Crouch, J.H., Madsen, S., Vuylsteke, D.R., Ortız, R., 2000. Comparative analysis of phenotypic andgenotypic diversity among plantain landraces (Musa spp., AAB group). Theor. Appl. Genet, 101:1056–1065 Dellaporta, S.L., Wood, J., Hicks, J.B., 1983. A plant DNA mini preparation: Version II. Plant Mol. Bio. Rep. 1: 19-21. Doyle, J.J., Doyle, J.L., 1990. Isolation of plant DNA from fresh tissue. Focus 12: 13-15. Edwards, K., Johnstone, C.,Thompson. C., 1991. A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Res. 19: 1349. Emberger, L., Maire, R., 1941. Cataloguedes plantes du Maroc.Volume IV. Minerva, Alger. Garcia, M. G., . Ontivero, M., J. Dıazrıccı, C., Castagnaro, A., 2002. Morphological traits and high resolution RAPD markers for the identification of the main strawberry varieties cultivated in Argentina. Plant Breeding 121, 76-80. Gharnit, N., El Mtili, N., Ennabili, A., Sayah, F., 2004. Floral characterization of carob tree (Ceratonia siliqua L.) from the province of Chefchaouen (NW of Morocco). Moroccan J. Biol., 1pp. 41–5 Guillemaut, P., and Maréchal-Drouard, L., 1992. Isolation of plant DNA: a fast, inexpensive and reliable method. Plant Mol. Biol. Rep. 10: 60-65. Güzeldağ, G.,. Çolak, Õ., 2007. Molecular identification of Ganoderma lucidm from Turkey. Int. J. Agric. Biol., 9: 767–770. Halliwell, B., 1990. How to characterize a biological antioxidant. Free Radic. Res. Commun. 9: 1-32. Jobes, D.V., Hurley, D.L., Thien, L.B., 1995. Plant DNA isolation: a method to efficiently remove polyphenolics, polysaccharides and RNA. Taxon 44: 349-386. Konate, I., Berraho, E.B., Filali-Maltouf, A., 2009. Inter-simple sequence repeat markers variation among natural accessions of Moroccan Carob Tree (Ceratonia siliqua L.), Inter. J. of Agri.Biol., 11: 168-172. Konaté, I., Filali-Maltouf, A., Berraho, E., 2007. Diversity analysis of Moroccan carob (Ceratonia siliqua L.) accessions using phenotypic traits and RAPD markers, Acta Bot. Malacitana, 32: 79–90. Kulkarni M, Borse, T., Chaphalkar, S., 2001.Isolation and purification of genomic DNA from black plum (Eugenia jambolana Lam.) for analytical applications. Int. J. Biotechnol. Biochem. 3: 49-55. Lodhi, M.A., Ye, G.N., Weeden, N.F., Reisch, B.I., 1994. A simple and efficient method DNA extraction from grapevine caltivars and vitis species. Plant Mol. Biol. Rep., 12: 6–13. Morsy, A.A., 2007. Molecular variation of Achillea fragrantissima (Forssk.) SCH. BIP. Growing in five areas of South Sinai. Int. J. Agric. Biol., 9: 11–17 Ounzar, B., Hartmann, C., Rode, A., Benslimane, A., 1998. Date palm DNA minipreparation without liquid nitrogen. Plant Mol. Rep. 16: 263-269. Samal S., Rout, G.R., Lenkaet, P.C., -2003- Analysis of genetic relationships between populations of cashew (Anacardium occidentale L.) by using morphological characterisation and RAPD markers. Plant Soil Environ. 49, (4): 176-182. Samarakoon, T., Wang, S.Y., Alford, M.H., 2013. Enhancing PCR amplification of DNA from recalcitrant plant specimens using a trehalose-based additive. Appl. Plant Sci. 1: 1200236. Sytsma, K., Givnish, T.J., Simt, J.F., Hahn, W.J., 1993. Collection and Storage of Land Plant Samples for Macromolecular Comparisons. In: Methods in Enzymology - Molecular Evolution: Producing the Biochemical Data (Zimmer EA, White TJ, Cann RL and Wilson AC, eds.). Academic Press, San Diego, 23-38. Talhinhas, P., Neves-Martins, J., Leilâo, J., 2003. AFLP, ISSR and RAPD markers reveal high levels of genetic diversity among Lupinus spp. Plant Breed., 122: 507–510. Tous, J., Batlle, I., 1990. In: El Algarrobo (Ed). Mundi-Prensa, Madrid. Tous,J., 1992. Isozyme polymorphism in carob cultivars Hort. Sci., 27 (1992), pp. 257–258 Weishing, K., Nybom, H., Wolff, K., Meyer, W., 1995. DNA isolation and purification. In: DNA fingerprinting in plants and fungi, pp 44-59. CRC Press, Boca Raton, Florida.
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Details
| Journal Section | ARAŞTIRMALAR |
|---|---|
| Authors | |
| Publication Date | October 14, 2017 |
| Published in Issue | Year 2017 Volume: 48 Issue: 2 |
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